Phospho-Raf-1 (S642) Antibody

Catalog #: PPS061 Datasheet / COA / SDS

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PPS061

All Raf-1 Antibodies

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Phospho-Raf-1 (S642) Antibody Summary

Species Reactivity

Bovine, Canine, Chicken, Primate

Specificity

Human, mouse, and rat ~73 kDa Raf-1 phosphorylated at S642 in Western blots.

Source

Polyclonal Rabbit IgG

Purification

Antigen Affinity-purified

Immunogen

Phosphopeptide corresponding to amino acid residues surrounding the phospho-S642 of Raf-1

Formulation

100 μL in 10 mM HEPES (pH 7.5), 150 mM NaCl, 100 μg/mL BSA and 50% glycerol.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

1:1000 dilution

UV treated human Jurkat cell lysates

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Reconstitution Calculator

Preparation and Storage

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

For long-term storage, ≤ -20° C is recommended. Product is stable at ≤ -20° C for at least 1 year.

Background: Raf-1

Raf-1 (also C-Raf) is a ubiquitously expressed, 73 kDa, 648 amino acid (aa) Raf subfamily, Ser/Thr protein kinase family enzyme. Raf-1 is an entry point into the MAP kinase/Erk-1/2 signaling pathway. The molecule contains three distinct regions; an N-terminal RBD (Ras-binding domain) (amino acids 56-131), followed by two rich segments [a cysteine-finger region (amino acids 138-184) (also called CR1/C1) and a second cysteine-rich region (CR2) (amino acids 253-264)] and a C-terminal Ser/Thr kinase catalytic domain (amino acids 354-611). In general, Raf-1 phosphorylates and activates MEK-1/2. MEK-1/2, in turn, phosphorylates and activates
ERK-1/2. The activation of Raf-1 begins upstream at the plasma membrane with the activation of Ras. During Ras activation, Raf-1 is inactive, in part because a
14-3-3 protein constitutively binds to phosphorylated Ser 259 and Ser 621 residues on Raf-1. Once activated, Ras binds Raf-1 on the Raf RBD, and displaces 14-3-3, allowing for additional Ras binding to the CR1. These two ligations now activate Raf-1. In the inactive state, Raf-1 is constitutively phosphorylated on Ser 43, Ser 259, and Ser 621. Upon activation, Ser 338, Tyr 431, Tyr 491, and Ser 494 also become phosphorylated, with phosphorylation at Ser 338 inducing activation. Down-regulation of Raf-1 occurs on other sites. Phosphorylation on proline-directed serine sites at position 301 and 642 are posited to be part of a negative feedback system regulated by ERK.

References

Wellbrock, C. et al. (2004) Nat. Rev. Mol. Cell. Biol. 5:875.
Dhillon, A.S. and W. Kolch (2002) Arch. Biochem. Biophys. 404:3.
Muslin, A.J. (2005) Trends Cardiovasc. Med. 15:225.
Morrison, D.K. and R.E. Cutler (1997) Curr. Opin. Cell Biol. 9:174.
Bonner, T.I. (1986) Nucleic Acids Res. 14:1009.
Dougherty, M.K. et al. (2005) Mol. Cell 17:215.
Hekman, M. et al. (2005) FEBS Lett. 579:464.

Long Name

v-raf-1Murine Leukemia Viral Oncogene Homolog 1

Entrez Gene IDs

5894 (Human); 110157 (Mouse); 24703 (Rat)

Alternate Names

cRaf; c-Raf; c-Raf-1; CRAFraf proto-oncogene serine/threonine protein kinase; EC 2.7.11; EC 2.7.11.1; NS5; Proto-oncogene c-RAF; RAF proto-oncogene serine/threonine-protein kinase; RAF; Raf1; Raf-1; Raf-1Oncogene RAF1; v-raf-1 murine leukemia viral oncogene homolog 1; v-Raf-1